1, 5-pregnadienes and processes for their manufacture



1,'5'-PREGNADIENES AND PROCESSES FOR THEIR MANUFACTURE Alexander L.Nussbaurn, Leonia, and Eugene Oliveto, Glen Ridge, N.J., assignors toSchering Corporation, Bloomfield, N.J., a corporation of New Jersey NoDrawing. Application February 2, 1959 Serial No. 790,380

29 Claims. (Cl. 260-39745) This invention pertains to a new group oftherapeutically active pregnadienes and to methods for theirmanufacture. In particular, our invention relates to1,5-pregnadiene-17a,21-diol-3,20-diones which are potentantiinflammatory agents.

Our novel compounds may berepresented by the following formula:

wherein X is a member of the group consisting of hydrogen and halogenhaving an atomic weight less than 126; Y is a member of the groupconsisting of O and (H, ,BOH); Z is a member of the group consisting ofhydrogen and an alkyl radical preferably having from 1 to 4 carbonatoms; and R is a member of the group consisting of H and a'cyl.

Illustrative of the 2-esters contemplated above are alkanoates such asacetate, propionate, tertiary butylacetate, cyclopentylpropionate,dimethylacetate, trimethylacetate and phenoXyacet-ate; aryl esters suchas benzoate, thiophene carboxylate, nicotinate; esters from dibasicorganic esters such as the succinate, phthalate, and the sulfobenzoate;and those from polybasic inorganic acids such as sulfate and phosphate.

Our novel compounds are prepared by subjecting a 1,4- pregnadiene or al6-alkyl-l,4-pregnadiene possessing a halogen substituent greater thanfluorine at -6 to the action of zinc in alcohol or magnesium in ether ortetrahydrofuran. By proper control of acidity and solvent polarity, the1,5-pregnadiene compounds are formed and isolated. Alternatively, thesubstituent at C-6 may be a pseudohalogen such as tosylate or mesylateor a lower alkanoyloxy group like acetoxy or propionoxy. When thereactive group in the 6-position is halogen or tosylate, the reactiontowards formation of the 1,5-dienes may proceed smoothly at roomtemperature. With a mesylate or lower alkanoyl ester in the 6-position,higher temperatures in the range of 50-100 C. are preferred to effectthe transformation since at lower temperatures the reaction proceedsquite slowly.

The C-6-substituted intermediates are prepared from3-keto-1,4-pregnadiene starting compounds in a variety of ways. Startingcompounds having a hydrogen at C-16 such as prednisone and prednisoloneare well known in the art, 'and the 16-lower alkyl-1,4-pregnadienestarting compounds are preferably prepared as described in copendingapplication Serial No. 733,843 of Rausser et al., filed May 8, 1958.Introduction of the halogeno, pseudohalogeno or acyloxy substituent at0-6 iscarried out by Patented Oct. 13, 1959 any one of several methods.For example, allylic halogenation of a 3-keto-l,4-pregnadiene or a3-keto-16-alkyl- 1,4-pregnadiene with a reagent such asN-bromosuccinimide, N-bromoacetamide, N-chlorosuccinimide, bromine orchlorine in such solvents as dimethylsulfoxide or dioxane yields thecorresponding'6-halogeno-l,4-pregnadiene or6-halogeno-16-alkyl-l,4-pregnadiene. Esterification of a6-hydroxy-1,4-pregnadiene or a G-hydfoxy- 16-alkyl-1,4-pregnadiene canbe made to yield the tosylate or mesylate or lower alkanoateintermediates. These -hydroxy-lA-pregnadiene and 6-hydroXy-l6-alkyl-L4-pregnadiene starting compounds are prepared by subjecting a6-desoxy-l,4-pregnadiene or a 6-desoXy-16-alkyl- 1,4-pregnadiene to themicrobiological oxygenating action of an organism of the genusChaetomium according to the analogous procedure described in BelgianPatent No. 548,450.

Prior to introducing a substituent to the 6-position of a 1,4-diene soas to prepare the immediate precursor of the compounds'of our invention,we prefer to esterify any reactive hydroxyl groups which may be present,such as at 0-11 or C2l. Thus, for example, in the case of prednisone,the 21-esters are the preferred starting compounds. Likewise, inpreparing 6-bromo-l6a-methylprednisone for ultimate conversion into16a-methyl-1,5- pregnadiene-lluJ1-diol-3,11,20-trione, we prefer toemploy a 2l-ester of l6m-methylprednisone, such as the acetate, as thestarting substance. In those instances where there is a hydroxyl groupat (3-11 such as when'a prednisolone or a l6u-methylprednisolone analogis employed as a starting substance or an intermediate, the conditionsrequired for est'erification of the C-ll group of necessity esterifyboth the 1741- and Zl-hydroxy groups as well. Thus when preparing a6-halogenoprednisolone or a l6oz-methylprednisolone derivative for useas an immediate precursor of a compound of this invention, an11,17,2l-triester such as prednisolone triacetate ormethylprednisolone-l1,17,2l-triacetate is employed. It is only in theparticular instance where 'a halogenation step follows thatesterificatio'n of a C-11 hydroxyl group is advisable. However, if theintermediate being prepared is an ester such as 6-alkanoate or a6-mesylate or a 6- tosylate, then it is only necessary to protect thehydroxyl group at C2 l, since the esterifi'cation at C-6 can be carriedoutselectivelyin the presence of both an 11 and 17-hydroxyl group.

It is apparent that the 16-alkyl-1,5-pregnadiene obtained will ofnecessity contain the original ester protecof the substituent at C6 andprior to conversion to a 1,5-diene. This process is applicable only tothose methods wherein the intermediary compound contains a halogen atC-6. The saponification of the ester may be effected by means of astrong acid such as sulfonic, perchloric, or p-toluenesulfonic inaqueous alcohol. By way of example, 16a-methylprednisone is esterifiedto' yield l6otmethylprednisone Zl-acetate, whereupon bromine isintroduced at 0-6. The 6,-bromo-16a-methylprednisone Zl-acetate may besaponified to form 6-bromo-l6a-methylprednisone which is then convertedinto the ra -analog by reacting said 6-bromo-l6a-methylprednisone withzinc in alcohol yielding 1dot-methyl-1,5-pregnadiene-170:,21- diol-3 ,11,20-trione.

By means .of the rearrangement process of this inven- 3 tion,1,4-pregnadienes and 16-alkyl-1,4-pregnadienes substituted in positionsother than those indicated in general Formula I may also be converted tothe corresponding 1,5-pregnadienes. Thus, 6a-methylprednisone 2l-acetateupon treatment with N-bromosuccinimide forms the 6-bromo intermediate.This latter compound when reacted with zinc in alcohol, yields6a-methyl-l7u,2l-dihydroxy-l,5-pregnadiene-3,l1,20-trione 2l-acetate.1,5- pregnadienes may also be prepared from other substituted1,4-pregnadienes such as 6a-methylprednisolone,6amethyl-9a-fluoroprednisolone, Z-methylprednisolone, 90c,2l-difluoro-2l-desoxyprednisolone, 16rx-hydroxyprednisolone and16a-hydroxy-9a-fluoroprednisolone in an analogous manner.

The process of our invention whereby a 4,5-double bond rearranges to a5,6-double bond is applicable not only to 1,4-pregnadienes but to anysteroid containing a 3-keto-A -configuration. Thus any pregnane,androstane, saponin, sapogenin or bile acid containing a 3-keto-A moietywhich will form a 6-substituted derivative such as the 6-halogeno-,6-tosylate-, 6-mesylate, or 6-acy1ate-, a requisite intermediarystarting compound, will form the corresponding 3-keto-A -product whensaid 6-substituted intermediate is treated with an agent such as zinc inethanol. Derivatives of 4-pregnenes such as cortisone, hydrocortisone,progesterone, l9-norprogesterone as well as ll-dehydroprogesterone, andthe like, are thus transformed to their corresponding A -pregnenes.Androstanes such as A -androstadiene-17fiol-3-one, testosterone, A

androstadiene-3,17dione, A -androstene, and the like are converted tothe A -androstenes.

The new 1,5-pregnadienes and 16-a1kyl-1,5-pregnadienes of our inventionfalling within Formula I are antiinflammatory agents useful inalleviating conditions such as rheumatoid arthritis, intractable asthma,rheumatic fever, disseminate lupus erythematosus; skin ailments such asextensive atopic dermatoses, as well as certain inflammatory disordersof the eye. Our 1,5-pregnadienes advantageously do not possess manyundesirable side eflfects which often accompany the use of corticalhormones. For example, l,S-pregnadiene-17a,21-diol- 3,11,20-trione is 3to 4 times as active as cortisone as an anti-inflammatory agent inalleviating arthritis, said diene having potency in the order of thatpossessed by prednisone. The 1,5-diene, when administered to a patienteven at twice the effective dose of prednisone, advantageously exhibitsless nitrogen, calcium and phosphorous loss, less sodium and waterretention, as well as less increase in fasting blood sugar as occurswith prednisone administration. We also have discovered that theintroduction of the l6-alkyl group into a 1,5-diene completelyeliminates the residual salt and water retaining properties of thesubstance, while simultaneously retaining the other advantageousproperties, i.e. lower nitrogen, calcium and phosphorous loss and lessincreasein fasting blood sugar. Thus, since our novel compounds exhibitlittle or no water or sodium retention, they may be taken in large dosesif necessary, providing for anti-inflammatory action without beingconcerned with edematous etfect.

Our therapeutically valuable compounds are preferably administeredorally in the form of tablets containing, for example about 2 to 25 mg.per tablet mixed with a solid carrier containing one or more of theusual exciplents such as starch, sugar, gums, soaps, clays and the like.Where parenteral administration is indicated, subcutaneous orintramuscular injection of a 2l-lower alkanoyl ester dissolved orsuspended in a suitable nontoxic liquid vehicle is preferred. In thetreatment of skin conditions such as atopic dermatoses, topicalpreparations such as containing 25% of active ingredient areadvantageously employed. In some instances, such as direct treatment ofan inflammed joint, crystalline suspensions are injectedintra-articularly. These microcrystalline suspensions are adaptable foruse in nasal sprays as well.

1 In addition to being active anti-inflammutQliQS, themselves, the newcompounds of our invention are useful intermediates in the production ofnew classes of physiologically active corticoids, indicated below asIII, IV, and V, these latter compounds being potent long actingantiinflammatory agents devoid of side efiects such as gastricirritation, salt and water retention and moon face.

Methods of utilizing our compounds as intermediates are illustrated bythe reactions outlined below wherein X, Y, R and Z have the meaningspreviously given.

CHzOR X\ H3C Peracid CH CH CHaOR CIIHnOR I NOR dCIIOR Y Y M Z W 11 0 moHydrohalic acid 0:

E E l O HO Halogen II III (E z (IJHzOR i i Y\ NOR .u-OR X MN Z X\ NW ZH,o\ 11,0

Acetic acid l l HO F F IIIA IV CH C 3 (IJHzOR (IIH OR 5 3 Y\ I'IIOR "NORX W Z X M Z 11,0 H O Zinc m Ethanol O O i 5 Ho Br on IIB V Epoxidationof the 3-keto-A -dienes with peracids such as peracetic, perbenzoic, andthe like, produces 5a,6aepoxy-3-keto-A -steroids II) which, on treatmentwith hydrohalic acids, yields A -3-keto-5a-hydroxy-6p-halosteroids asexemplified by III. Dehydration of 1-H with acetic acid, for example,produces 6fl-fluoro-A -dienc-3- ones ('IV). Dehalogenation of theGfi-bromo derivatives, I-I I, with a reactant such as zinc in ethanol,chromous chloride, or Raney nickel gives 5a-hydroxy-A -3-keto steroids(V).

Similarly, any A -3-keto steroids may undergo the above indicatedtransformations to form derivatives analogous Some of thephysiologically active compounds which have been made as indicated aboveare as follows. SKI-hydroxy-6-halogeno compounds of the type representedby Formula Il l are 613 fluoro 5u,17a,21 trihydroxy l pregnene 3,11,20trione, 6B fluoro 5a,11}3,17oc,21 tetrahydroxy l pregnene 3,20 dione,613,90: difluoro- 5u,17a,21 trihydroxy 1 pregnene 3,11,20 trione, 613,9difluoro 5ot',11,8,-17a,21 tetrahydroxy l pregnene 3,20 dione and 6Bfluoro 6a methyl 5a,-l7a,21 e trihydroxy 1 pregnene 3,11,20- trione andthe 16-methyl analogs of the foregoing.

Active corticosteroids of the type-represented by Formula :IV are6p-fluoroprednisone and '21-acetate, 6,8- fiuoroprednisolone and21-acetate, 6fi,9a-difluoroprednisome and 2lacetate,6B,9u-difluoroprednisolone and 21- acetate,6a-methyl-6B-fiuor0prednisone and Zl-acetate,6a-methyl-6,8-fluoroprednisolone and 21-acetate, 613-fluoro-l6-methylprednisones and their 21-acetates, 65-fluoro-16-methylpredniso1ones and their 21-acetates,6,8,9a-difiuoro-16a-hydroxyprednisone and 16,21-diacetate, and6fi,9u-difluoro-loa-hydroxyprednisolone and 16,21-diacetate.

Some valuable compounds of the type represented by Formula V are5a,17oc,21 trihydroxy 1 pregnene 3,11,20 trione, 5a,1l/3,l7u,2ltetrahydroxy 1 pregnene 3,20 dione, 90c fluoro 5oc,17ot,21 trihydroxy 1pregnene 3,11,20 trione, 9oz fluoro 5a,11}8,17ot,21 tetrahydroxy 1pregnene 3,20 dione, 6a methyl 50,17ot,21 trihydr oxy 1 pregnene 3,11,20trione, 6a methyl 5a,11/3,l7a,21 tetrahydroxy 1 pregnene 3,20 dione, 16methyl 5a,17ot,21-- trihydroxy 1 pregnene 3,11,20 triones, 16 methyl5u,11,8,17a,- 21 tetrahydroxy 1 pregnene 3,20 diones, 9a fluoro5a,l6ot,17ct,2l tetrahydroxy 1 pregnene 3,11,20 trione, and 90a fluoro5a,11[3,16a,l7u,21- pentahydroxy 1 pregnene 3,20 dione.

All the compounds of our invention are valuable therapeutic agents asoutlined above. The preferred embodiment of our invention, however, arethe 9a-fluoro analogs of the 1,5-pregnadienes falling within Formula I,and particularly the 9a-fluoro analogs of the 16-methyl-1,5-pregnadienes.

The present invention is a continuation-in-part of our co-pendingapplications Serial No. 725,521, filed April 1, 1958, and Serial No.770,315, filed October 29, 1958.

The following examples are illustrative of the procedure employed inpreparing the compounds of this invention, but are not to be construedas limiting the scope thereof; the scope of our invention being limitedonly by the appended claims.

EXAMPLE 1 17m,21-dihydroxy-1,5-pregnadierte-3J1,20- trione 21 -aceiafeA. 6,8 bromo 17oc,21 dihydrox y- 1,4 pregnadiene 3,11,20-trione21-acetate (500 mg.) is dissolved in 150 ml. of absolute alcohol, then25 ml. of water and 5 g. of zinc powder are added. The suspension is.stirred at room temperature for 8 hours, after which time the zinc isfiltered, and the filtrate concentrated to dryness in vacuo. Theresulting residue is recrystallized from ace tone to yield 312 mg. ofl7a,2l-dihydroxy-l,5-pregnadiene-3,11,20-trione 21-a'cetate, M.P.214-218 C.,

Analysis.Calcd. for C26H34O7I C, 68.10; H, 7.47. Found: C, 67.87; H,7.39.

B. Alternatively, the compound of this example may be prepared asfollows. 6B,l7a,2l-trihydroxy-1,4-pregnadiene-3,1l,20-trione6,2l-diacetate (500 mg.) is dissolved in 150 ml. of absolute ethylalcohol, then 25 ml. of water and 5 g. of zinc powder are added. Thesuspension is stirred for one hour at 90 C., after which the solution iscooled, the zinc filtered, and the filtrate 6 concentrated to dryness invacuo. The resulting residue is recrystallized from acetone to yield17a,21'-dihydroxy- 1,5 pregnadiene 3,11,20 trione ZI-acetate, M.P. 214-218 C.

EXAMPLE 2 1 7OL,21 -dihydroxy-1,5-pregnadiene-3,11,20-tri0ne The1,5-pregadiene 21-acetate prepared in Example 1 is hydrolyzed to thecorresponding 21-hydroxy compound with the aid of a culture of Flavobacterium dehydro genans (Rutgers University Collection No.

A. The culture of the organism is prepared by propagating it in anutrient agar medium at 30 C. [for 24 to 72 hours. During incubation,the inoculated tube is exposed to light with the resultant developmentof a yellow pigment characteristic of the species. The developed cultureis rinsed from an agar slant under sterile conditions into a sterilemedium of pH 6.8 and having the following composition:

Gm. Yeast extract (Difco) 10 Potassium phosphate monobasic 4.48 Sodiumphosphate dibasic 4.68

Tap water to 1 liter.

This culture medium has previously been autoclaved, as at 15 lbs.pressure, for twenty minutes toobtain aseptic conditions, and cooled.The variant is grown in are introduced into each of ten flasks, andt'oeach'flask are added 200 mg. of 17o,2l-dihydroxy-1,5-pregnadiehe-3,11,20-trione 2l-acetate dissolved in a minimum volume of ethanol. Thereaction mixtures are then shaken at 30 C. for 12 to 72 hours. Thereaction is stopped when paper chromatography indicates that thestarting material has been transformed.

The contents of the flasks are combined and extracted with methylenechloride. The extracts are concentrated and the residue is crystallizedfrom acetone-hexane afio'rding 0.62 g. of17ot,21-dihydroxy-1,5-pregnadiene-3,11,20- trione as a crystallinesolid.

Alternatively, the compound of this example may be prepared by thefollowing procedures B and C.

B. 613 bromo ;,21 dihydroxy 1,4 pregnadiene 5,11,20-trione.'6B bronio1704,21 dihydroxy-lA-p'regnadiene-3,ll,20-trione 2l-acetate (500 mg.) isdissolved in 25 ml. of absolute methanol and 5 of water.p-Toluenesulfonic acid (100 mg.) is added and the resulting reactionmixture allowed to stand overnight at room temperature. The alcohol isthen evaporated in' vacuo, water added to the residue, and the resultingprecipitate filtered and dried at room temperature. Recrystallization ofthis solid from acetone-hexane gvies 6fl-bromo-170:,21-dihydroxy-1,4-pregnadiene-3 ,1 1,20-trione.

C. 17a,21-dihydroxy-1,S-Pregnadiene-SJ1,20-tri0ne. 6B bromo- 170:,21:dihydroXy-1,4 pregnadiene 3,11,20- trione (500 mg), prepared as in aboveExample 2B, is reacted with zinc in ethanol and water in the manner ofExample 1A to yield 17,21-dihydroxy-l,5-pregnadiene 3,1 1,20-trione.

EXAMPLE 3 3,-l1,20-trione 21-acetate is reacted with zinc, ethanol andwater in the manner described in Example 1A, to yield9a-fiuoro-l7a,2l-dihydroxy- 1,5 -pregnadiene-3, 1 1,20-

trione 21-acetate which is crystallized from ethyl acetate.

Alternatively, the compound of this sample may be prepared as follows:

6}8-bromo-9a-fluoro-17a,21-dihydroxy-1,4-pregnadiene- 3,11,20-trione21-acetate (500 mg.) is dissolved in 10 ml. of tetrahydrofuran, and 4 g.of magnesium turnings added. After refluxing this reaction mixture forone hour, it is cooled to room temperature, and 25 ml. of a 5% aqueousammonium sulfate solution added. After stirring this mixture an hour,chloroform is added, and the organic solvent layer separated from theaqueous layer. The chloroform solution is concentrated in vacuo to yield9afluoro- 170:,21- dihydroxy-l,5- pregnadiene- 3,11,20- trione21-acetate.

In similar fashion, other new compounds of this invention as representedby Examples 19 may be prepared using magnesium in an inert solvent suchas ether or tetrahydrofuran.

The term inert solvent as used in this application is defined as anorganic solvent which itself does not react or enter into the reactionsdescribed.

EXAMPLE 4 9ot-flur0r0-1 7a,21-dihydr0xy-1 ,5 -pregnadiene-3,11,20-trione The 9a-fluoro-1,5-pregnadiene 21-acetate of Example 3 issubjected to a culture of F lavobacterium dehydrogenans in the manner ofExample 2 to give 9a-fluoro-17u,21- dihydroxy-1,5 -pregnadiene-3,11,20-trione.

EXAMPLE 5 115,170:,21-trihydrxy-1,5-pregnadiene-3,20-di0ne A.6fl-br0m0-115,I7a,21-trihydr0xy 1,4 pregnadz'ene- 3,20-dionetriacetate.To 1113,17a,21-trihydroxy-1,4-pregnadiene-3,20-dionetriacetate (5.2 g.) dissolved in 1500 ml. of carbon tetrachloride thereis added 2.53 g. of N- bromosuccinimide and 0.075 g. of benzoylperoxide, and the mixture is refluxed with illumination (immersed bulbRFL No. 2) for 30 minutes. After slightly cooling the reaction mixture,water is added and the organic layer separated and concentrated to anoily residue of 6B- bromo-11fl,17u,21-trihydroxy-1 ,4-pregnadienetriacetate used Without further purification in the following reaction.1

B. 115,] 70:,21- trihydroxy- 1,5- pregnadiene-3,20- dionetriacetate.--6/3-br0m0-l 1B,17oz,2l trihydroxy-1,4-pregnadiene-3,20-dione triacete (200 mg.) is dissolved in 70 ml. ofabsolute methyl alcohol then 10 ml. of Water and 2 g. of zinc powder areadded. After the suspension is stirred at room temperature for sevenhours, the zinc is filtered, and'the filtrate concentrated to acrystalline residue which is recrystallized from ether to yield11B,l7a,21-trihydroxy-1,5-pregnadiene-3,20-dione triacetate, M.P.155-160 C.,

C. I1fl,17a,21 trihydroxy 1,5 pregnadiene 3,20-dione.-The'1,S-pregnadiene-triacetate prepared in above Example B issubjected to the action of a culture of F lavobacterium dehydrogenans inthe manner of Example 2. to yield11B,170:,21-trihydroxy-1,5-pregnadiene-3,20- dione which may berecrystallized from methanol.

Alternatively, the compound of this example may be prepared as describedin following procedures D and E.

D. 6fi,]1fl,17ot,21 tetrahydroxy 1,4 pregnadienc- 3,20-di0ne6-methanesulfonate 2I-acetate.100 mg. of 6,3,11fl,17a,21 tetrahydroxy1,4 pregnadiene 3,20- dione 21-acetate (prepared as described in BelgiumPatent No. 548,450) is dissolved in 0.4 ml. of methylene chloride and0.4 ml. of pyridine. The solution is cooled to 0 C., 1.6 ml. ofmethanesulfonyl chloride added, and the mixture stirred for 2 hours at 0C., then left overnight at room temperature. The reaction mixture is 8then poured into water, the resulting precipitate which is filtered andair dried is 65,1lfl,17oz,21-tetrahydroxy-1,4- pregnadiene-3,20-dioneG-methanesulfonate 21-acetate.

E. 11B,17a,21 trihydroxy 1,5 pregnadiene 3,20- aiona-The 6methanesulfonate 1,4 pregnadiene (55 mg.) of above Example 5D isdissolved in 20 ml. of ethanol and'l ml. of water. Zinc filings (100mg.) are added, and the suspension stirred for 1 hour at C. The solutionis cooled, the zinc filtered, and the resulting filtrate concentrated toa residue substantially of 115,170,21-trihydroxy-1,5-pregnadiene-3,20-dione 21-acetate.

This 21-acetate residue is hydrolyzed in the manner of Example SC toyield 1lfl,17a,21-trihydroxy-1,5 pregnadiene-3,20-dione.

A third alternative procedure of preparing the compound of this exampleis given below.

F. 11,8,17a,21 trihydroxy 1,5- pregnadiene 3,20- dione 21acetate.6/3,l113,1711,21-tetrahydroxy-1,4-pregnadiene-3,20-dione6-p-toluenesulfonate 21-acetate (Belgium Patent No. 548,450) as astarting compound is reacted with zinc powder at room temperature in themanner described in Example 1A to obtain l1fi,17u,2ltrihydroxy-1,5-pregnadiene-3,20-dione 21-acetate.

This 21-acetate is hydrolyzed in the manner of Example 2 to yield1lfl,17a,21-trihydroxy-1,5-pregnadiene- 3,20-dione.

A fourth alternative for the preparation of this example is described inprocedures G and H.

G. 613 bromo 116,17a,21 trihydroxy 1,4 pregnadiene 3,20 dione-65 bromo11B,17a,21 trihydroxy-1,4-pregnadiene triacetate prepared as in Example5A, is subjected to the action of a culture of Flavobacteriumdehydrogenans in the manner of Example 2 to yield6B-bromo-11B,17a,21-trihydroxy-1,4-pregnadiene-3,20-dione.

H. 11fi,17a,21 trihydroxy 1,5 pregnadiene 3,20- di0ne.The6fi-bromo-11,8,170:,21-trihydroxy-1,4-pregnadiene-3,20-dione of aboveExample 56 is reacted with zinc and propyl alcohol in the manner ofExample 1A to yield 11B,17a,21 trihydroxy 1,5 pregnadiene 3,20- dione.

A fifth alternative for the preparation of this example is described inthe following procedure I.

I. 6 3,11B,17u,21 tetrahydroxy 1,4 pregnddiene-3,20-dione.6,2l-diacetate (prepared as described in Belgium Patent No.548,450) is reacted with zinc in ethanol in the manner described inExample 1A to yield 1lfl,17a,21 trihydroxy-1,5-pregnadiene 3,20 dione21- acetate.

A sixth alternative procedure is described in procedures J and K.

J. 6/3 bromo 115,1 7a,21 trihydroxy 1,4 pregnadz'ene 3,20 dione 21acetate-1 l,8,17a,21' trihydroxyl,4-pregnadiene-3,20-dione 21-acetate (1g.) is suspended in 10 ml. of dioxane and 5 ml. of acetic acid. To this,bromine (2.5 g.) dissolved in acetic acid (5 ml.) is added, and theresulting orange solution allowed to stand at room temperature for onehour. About ml. of ice-water is then added. The resulting precipitate of6/3-bromo- 11fi,17a,21 trihydroxy 1,4 pregnadiene 3,20 dione 21-acetateSKEHOH) 9 trihydroxy-I,5-pregnadiene-3,2O dione 21 acetate, M.P. 219-224C.,

EXAMPLE 6 Qa-fluoro-JIBJ7a,21-trihydr0xy-1,5-pregnadiene-3,20-- dione 65bromo 9a fluoro 11,8,l7a,21 trihydroxy 1,5- pregnadiene-3,20-dionetriacetate is treated with zinc powder and methyl alcohol in the mannerof Example B to give9a-fluoro-11/3,17a,21-trihydroxy-1,5-pregnadiene-3,20-dione triacetate.

The above triacetate of this example is subjected to the action of aculture of F lavorbacterium dehydrogenans in the manner of Example 2 toyield 90t-fiuOI'O-11fl,170t,21- trihydroxy-1,5-pregnadiene-3,20-dione.

One gram of the trihydroxy-1,5-pregnadiene of this example is dissolvedin 5 ml. of acetic anhydride, kept at 40 C. for 2 hours, and then pouredinto ice water. The resulting precipitate is filtered, air dried, andrecrystallized from methanol to yield9a-fiuoro-llfi,l7a,2l-trihydroxy-l,S-pregnadiene-S,20-dione 21-acetate.

EXAMPLE 7 9ct-br0mo-1 7oc-21 -dihydr0xy-1 ,5 -pregnadiene-3,1 1 ,20-trione 21-acetate A. 6,8,90: dibromo 170;,21 dihydroxy 1,4 pregnadiene3,11,20 trione 21 acetate-90: bromo 17a, 21-dihydroxy-1,4-pregnadiene3,11,20 trione 2l-acetate (1 g.) is dissolved in 200 ml. ofchlorobenzene and 225 ml. of carbon tetrachloride, then 25 ml. ofsolvent distilled under argon to remove any traces of moisture. N-bromosuccinimide (0.53 g.) is added, and the solution stronglyilluminated with an immersed bulb REL No. 2 while the solution is heatedand refluxed for 15 minutes. The reaction mixture is then cooled, pouredinto methylene chloride and Washed with water. The organic layer isdried over magnesium-sulfate, filtered, and concentrated to an oilyresidue of 65,9a-dibromo-17a,21-dihydroxy-1,4-pregnadiene-3,11,20-trione 21-acetate. The oily 6-bromide thus. obtained, is usedwithout further purification in the subsequent reaction.

B. 90: bromo 170:,2] dihydroxy 1,5 -pregnadiene- 3,11,20-tri0ne21-acetaze.The oily 6-bromide of above Example 7A. is reacted with zincin ethanol in the manner of Example 1. The product, isolated iscrystallized from. acetone to yield9oc-bromo-1704,21-dihydroxy-1,5-pregnadiene-3,11,20-trione ZI-acetate.

By subjecting the 21-acetate of this example to the action of Flavorbacterizmz dehydrogenaizs as described in Example 2 there isobtained 9a-bromo-l7oc,2l-dihydroxy- 1,5 -pregnadiene -3,1 1,20-trione.

EXAMPLE 8 1701,21 -dihydr0xy-1,5-pregnadiene-3,11,20-tri0ne 21-isobutyrate A. 6,8 bromo 17u,21 dihydroxy 1,4 pregnadiene- 3,11,20trione 21 is0butymte.17a,21 dihydroxyl,4-pregnadiene-3,11,20-trione21-isobutyrate (1.1) is reacted with N-bromosuccinimide (0.6 g.) in themanner of Example 7A to yield 6/3-bro1no-17a,21-dihydroxy-1,4-

an oily 1 0 EXAMPLE 9 9u-chl0r0-1:,21-dihydroxy-1,5-pregnadiene-3,11,20- trione9a-chloro-17a,2'1-dihydroxy -1,4 pregnadiene 3,11,20 trione 21-acetateis reacted with N-brornosuccinimide inthe' manner of Example 7A to yield6fi-bromo-9a-chloro- 17a,21-dihydroxy-1,4-pregnadiene-3,11,20-trione21-acetate as an oily residue.

This oily 6,8-bromide is reacted with zinc in ethanol in the manner ofExample 1A to give c-Cl1l0rO-l7cc,21 dihydroxy-l,5-pregnadiene-3,11,20-trioue 21-acetate.

This 21-acetate is hydrolyzed to the corresponding 21-alcohol in themanner of Example 2 to yield 9a-chloro-17a,21-dihydroxy-1,5pregnadiene-3,11,20-trione.

EXAMPLE 10 A. 6B-br0m0-9ot fluoro-16a methyl-11fl,17a,21trihydroxy-I,4-pregnadiene-3,20-di0ne 21-acetate.-Therequisite starting material, 9a-fluoro-16a-methyl-l1fi,17a,21-trihydroxy-l,4-pregnadiene-3,20-dione 2l-acetate, is pre pared by theprocess described in co-pending application Serial No. 733,843 ofRausser et al., filed May 8, 1958.

9a fluoro 16a methyl 11B,17a,21 trihydroxy 1,4- pregnadiene-3,20-dione21-acetate (380 mg.) is dissolved in 45 ml. of dioxane and then 165 mg.of bromine in 20 ml. of dioxane is added to the stirred solution at roomtemperature. After the color is completely discharged, agitation iscontinued for another ten minutes before pouring the solution into 500ml. of ice-water. The precipitate which thus forms is filtered and airdried, and is substantially 6/3-bromo 9a-flu0ro-16a-methyl-11,8,17u,2l-trihydroxy-l,4-pregnadiene-3,20-dione 21-acetate. This product isused without further purification in the following procedure.

B. 9ot-flu0r0J6a-methyl 11,8,17ot,21 trihydroxy 1,5-pregnadiene-3,20-di0ne 21 -acetate.6B-bromo-9a-fiuoro- 16a methyl-l1B,170;,21-trihydroxy-1,4-pregnadiene-3,20- dione 21-acetate,prepared. in above Example 10A, is dissolved in ml. of ethanol to which15 ml. of water is added and the resulting solution is heated to refluxtemperature. Zinc powder (1.5 g.) is then added andthe refluxingsuspension is stirred for 1.5 hours, cooled and filtered. The filtrateis concentrated to dryness in vacuo, and chromatographed over 30 g. ofFlorisil. The fractions eluted with benzene-ether (3:1) give 9a-flll0l0-a methyl -11/3,17u,21-trihydroxy-1,5-pregnadiene-3,20 dione 21-acetate.

EXAMPLE 1'1 The 16u-methyl-1,5-pregnadiene 21-acetate prepared in aboveExample 10 is hydrolyzed to the corresponding 21-hydroxy compound. withthe aid of a culture of Flavobacterium dehydrogenans in the mannerdescribed in Example 2. The product is isolated and purified in thedescribed. manner to give Qa-flUOIO-l6a-meihYl-ll/3,17u,21-trihydroxy-1,5-pregnadiene-3 ,20- dione.

EXAMPLE 12 A. 6,8 bromo 9m fluoro 1613 methyl 11fl,]7oz,21 trihydroxy-1,4 pregnadiene-3,20-dione 21-acetate.The requisite intermediate,9a-fiuoro-16fi-methyl1173,17a,21- trihydroxy-l,4-pregnadiene3,20-dione21-acetate, is prepared by the process. described in co-pendingapplication Serial No. 733,843 of Rausser et al.,.filed May 8, 1958;

9a fluoro 1'65 methyl 11B,17a,21 -trihydroxy-1,4- pregnadiene-3,20-dioneZI-acetate (380 mg.) is brominated andthe resulting productisolated inthe manner described in. Example 10A to give 6fl-bromo-9a-fluoro 11 16 3methyl -11B,17a,2l-trihydroxy-1,4-pregnadiene-3,20- dioue 21-acetate.

B. 90: fluoro -16B- methyl -11,8,17a,21 trihydroxy-LS-pregnadiene-3,20-dione 21-acetate.The6}3-bromo-9afluoro-16fi-methyl-1,4-pregnadiene of above Example 12A isreacted with zinc powder is ethanol-water in the manner of. Example 10B,and the resulting product purified in the described manner to give9a-fluoro-16fi-methyl-11fi,170:,2l-trihydroxy-1,5-pregnadiene-3,20-dione 21-acetate.

EXAMPLE 13 9a-fluor0-1 6fi-methyl-11/3J 7m,2 1-trihydroxy-1,5-pregnadiene-3,20-dione The 16fl-methyl-1,5-pregnadiene 21-acetate ofabove Example 12 is converted to the corresponding 21-alcohol with theaid of a culture of Flavobacterium dehydrogenans in the manner describedin Example 2 yielding 9a fluoro 16B-methyl -11fl,17a,21-trihydroxy-l,5-pregnadiene-3,20-dione.

Alternatively, the 16fi-methyl-11B,17a,21-trihydroxy- 1,5-pregnadiene ofthis example may be prepared as follows:

507 mg. of 6,8-bl'OlTlO-90t-flll0l'0-160L-methyl-11[3,170t,21-trihydroxy-l,4-pregnadiene-3,ZO-dione 21-acetate (prepared as in Example10A) is dissolved in 25 ml. of ice-cold ethanol containing 2.5 ml. ofwater and 65 mg. of sodium hydroxide. The solution is stirred undernitrogen at C. for 15 minutes, then neutralized with acetic acid andconcentrated in vacuo almost to dryness to yield a residue ofsubstantially 6B-br0mo-9u-fluoro- 1613 methyl-11B,170;,21-trihydroxy-1,4-pregnadiene-3,20- dione. To this 6fl-bromoresidue there is added ice-water and ethanol and the solution is reactedwith Zinc in the manner of Example B to give 9a-fluoro-16fl-methyl- 115,17a,21-trihydroxy-1,5-pregnadiene-3 ,20-dione.

EXAMPLE 14 9a-chlor0-1 6a-methyl-11B,17a,21-trihydroxy-1,5-pregnadiene-3,20-dione A. 6/3 bromo 9oz chloro -16o-methyl-11/3,17oz,Z1-tlihydroxy-I,4-pregnadiene-3,20-dione 21-acetate.Therequisite intermediate,9a-chloro-l6cx-methyl-116,17a-21-trihydroxy-l,4-pregnadiene-3,20-dioneZI-acetate, is prepared in the manner described in co-pendingapplication Serial No. 733,843, of Rausser et al., filed May 8, 1958.

90c chloro -16a-methyl-11/8,17a,21-trihydroxy1,4-pregnadiene-3,20-dione21-acetate is brominated, and the resulting product isolated, in themanner of Example 10A to give6,8-bromo-9u-chloro-16a-methyl-11B-17a,21-trihydroxy-l,4-pregnadiene-3,20-dione21-acetate.

B. 904 chloro -16a methyl -11{3,17a,21-trihydr0xy-1,5-pregnadiene-3,20-di0ne 21-acetate.-The 6/3-bromo-pregnadiene of aboveExample 14A is reacted with zinc and aqueous ethanol in the manner ofExample 10B, and the resulting product isolated to give9a-chloro-16a-methyl- 1l;8,17 x,2l-trihydroxy -1,5 pregnadiene 3,20dione 21- acetate.

C. 9a-chloro-16a-methyl 11B,17a,21 trihydr0xy-1,5-pregnadiene-3,20-dione.The 21-acetate of the 9u-ch1oropregnadieneprepared in above Example 14B is converted to the corresponding21-hydroxy with the aid of a culture of Flavobacterium dehydragenans inthe manner of Example 2 to yield9a-chloro-16ot-methyl-1lB,17a,21-trihydroxy-1,5-pregnadiene-3,20-dione.

Alternatively, the compound of this example may be prepared from6fi-bromo-9a-chloro-16a-methyl-11,3,17a,21-trihydroxy-1,4-pregnadiene3,20-dione 21-acetate (prepared as inExample 14A) in the manner described in the alternative procedure ofExample 13 whereby the 21- acetate of-the 6/3-bromo-1,4-pregnadiene isconverted to the corresponding 21-hydroxy which, in turn, is reactedwith zinc in ethanol-water to give 9ot-chloro-16a-methyl- 11,8,170:,21-trihydroxy-1,5-pregnadiene-3 ,ZO-dione.

. 12 EXAMPLE 15 9oc-chloro-16fl-methyl 11fi,17a,21trihydroxy-IJ-pregnadiene-3,20-di0ne A. 6/3-br0m0-9a-chlor0-16 3-methyl11fi,17a,21 trihydroxy-I,4-pregnadiene-3,20-di0ne 21-acetate.Therequisite intermediate,9a-chloro-16fl-methyl-11fl,17a,21-trihydroxy-l,4-pregnadiene-3,20-dione2l-acetate, is prepared according to the procedure described inco-pending application Serial No. 733,843 of Rausser et al., filed May8, 1958.

The 9a-chloro-16fl-methyl 11B,17oc,21 trihydroxy-1,4-pregnadiene-3,20-dione 21-acetate is brominated and the resultingproduct isolated in the manner of Example 10A to give 68-bromo-9u-chloro-16B-methyl-11p,l7a,21-trihydroxy-1,4-pregnadiene-3,20-dione21-acetate.

B. 9oz-chl0ro-16fi-methyl 11,8,17a,21 trihydroxy-1,5-pregnadiene-3,20-dione 21-acetate.The 6fl-bromo-9achloro-1,4-pregnadieneof above Example 15A is reacted with zinc powder in ethanol-water in themanner of Example lOB to give9m-chloro-16p-methyl-11B,17a,21-trihydroxy-l,5-pregnadiene-3,20-dione21-acetate.

C. 9a-chloro-16fi-methyl 11,8,17a,21 trihydroxy-1,5-pregnadiene-3,20-di0ne.The 21-acetate of the 9a-chloro-16fl-methylpregnadiene, prepared in above Example 15B, is converted tothe corresponding 21-hydroxy with the aid of a culture in F lavobacterz'um dehydrogenans in the manner of Example 2 to give9rx-Chl0I'O-l6/3-Il16tl1Yl-115,170,2l-trihydroxy-l,5-pregnadiene-3,20-dione.

Alternatively, the 21-alcohol of this example may be prepared by thealternative procedure of Example 13 whereby the6fi-bromo-9oc-chloro-16B-methyl-11fl,l7a,21- trihydroxy-l,S-pregnadiene3,20 dione 21-acetate, prepared in Example 15B, is hydrolyzed to thecorresponding 21-alc0ho1 which, in turn, is reacted with zinc inethanol-water to give 9a-chloro-16fi-methyl-11fl,17a,21-trihydroxy-1,5-pregnadiene-3,20-dione.

EXAMPLE 16 9a-flu0r0-16a-methyl :,21dihydroxy-LS-pregnadiene-3,11,20-tri0ne A.6,8-br0m0-9a-flu0ro-16oc-methyl 170;,21 dihydroxy-1,4-pregnadiene-3,11,ZO-trione 21-acetate.-The requisite intermediate,9u-fluoro-l6oc-methyl-17a,21-dihydroxy-1,4- pregnadiene3,l1,20-t-rione2l-acetate is prepared in the manner described in co-pending applicationSerial No. 733,843 of Rausser et al., filed May 8, 1958.

9ufluoro-16a-methyl 17,21 dihydroxy-l,4-pregnadiene-3,11,20-trione21-acetate is brominated, and the resulting .product isolated in themanner described in Example 10A to yield6,9-bromo-9a-fiuoro-l6a-methyl-17u,2l-dihydroxy-1,4-pregnadiene-3,11,20-trione21-acetate.

B. 9a-fluoro-16u-methyl 1711,21 dihydroxy-LS-pregnadiene-3,11,20-trione21-acetate.The 6;8-bromo-1,4- pregnadiene of above Example 16A isreacted with zinc powder in aqueous ethanol in the manner of Example 10Band isolated, and purified in the described manner to give9zx-fluoro-16ot-methyl-17a,2l-dihydroxy-1,5-pregnadiene-3,11,20-trione21-acetate.

C. 9a-flu0ro-16a-methyl 17u,21dihydroxy-LS-pregnadiene-3,11,20-trione.The 9oc-flll01'0 1,5 pregnadiene21-acetate of Example 16B is converted to the corresponding 2l-hydroxyby the action of a culture of Flavobacterium dehydrogenans in the mannerof Example 2 to yield 9a-fluoro-l6a-methyl-17a,2l-dihydroxy-1,5-pregnadiene-3 ,11,20-trione.

Alternatively, the compound of this example may be prepared in themanner described in the alternative procedure of Example 13, whereby6fl-bromo-9a-fiuoro-l6umethyl-1711,21-dihydroxy-1,4-pregnadiene 3,11,20trione 21-acetate (the compound of Example 16A) is first hydrolyzed tothe corresponding 21-alcoho1, then reacted with zinc to yield9a-fluoro-16u-methyl-flail-dihydroxy-' A. 6fi-bl'0m0 -9a-flll010 16,3methyl 170:,21 {dihydroxy-1 ,4-pregnadiene-3,1 1 ,ZO-trione 21 acet'ate.--'Ihe requisite intermediate,9ol-fluoro-16p-methyl-17ot,21-dihydroxy-l,4-pregnadiene-3,11,20-trioneZI-acetat-e, ispre; pared by the process described in co-pendingapplication Serial No. 733,843 of Rausser et al.; filed May 8, 1958.

In the manner of Example 10A, 9ot-fluoro l6fl-methyl-17u,2l-dihydroxy-1,4 pregnadiene-3,l1,20-trione 21-acetate is brominatedand the resulting product isolated to give6B-bromo-9ot-fluoro-16/3-methyl-l7 u,21 dihydroxy- 1,4-pregnadiene-3, 11,20-trione ZI-acetate.

B. 9a-fluoro-16fl-methyl 1704,21; dihydroxy-LS-pregnadiene-3,11,20-trl'0ne 21-aceltlte.-In the manner of EX- ample 10B, the6fi-bromo-L4-pregnadiene of above Example 17A is reacted with zinc inaqueous ethanol to give 9et-fluoro-16 8-methyl 1706,21dihydtoxy-1,5-pregnadiene-3,11,20-trine ZI-acetate.

Q. 9a-flllor0-16fl-methyl 170 21 dihydroxy- 1,5-pregnadiene-3,11,20-t'ibn.-Inthe manner of Example 2, the9ol-fluoro-16fl-methyl-l,5-pregnadiene 21 acetate of Exarn ple 17B, isconverted to the corresponding 2l-alcohol by the action of a culture ofF ldvobacterium de'hydrbgenans to give 9ot-fluoro-16fimethyl-17a,21-dihydroxy-1,5-pregnadiene-3,1 1,20-trione.

Alternatively, the compound of thisexample may be prepared by thealternate procedure of Example 13 from 6fi-bromo-9ot-fluoro-16,6-methyl17bt,21 dihydroxy-1,5-

pregnadiene-3,l1,20-trione 2l-acetate (the'c ompound of Example 17A) togive 9n -fluoro16p-methyl-17u,21-dihydroxy-1,5-pregnadiene-3 ,11,20-t'rione.

EXAM: PLE 18 A. 6/3,9oc-dibr0m0-16/3methyl 1113,170 21trihydroxy-1,4-pregn'adiene 3,20 dione 21 -acetate.-The requisiteintermediate, 9a-bromo-1'6'B-methyl-11;8,17a,21trihydroxy-1,4-pregnadiene-3,20-dione ZI-acetate, is prepared by theprocedure described in co-pending application Serial No. 733,843 ofRausser et al. filed May 8, 1958.

In the manner described in Example 10A, the 9w bromo-1,4-pregnadie'neintermediate of this example is brominated, and the resulting productisolated to give 6fi,9a-dibromo-16/3-methy1 11[3,l7a,21 trihydroxy-1,4-pregnadiene-3,20-dione 2 1-acetate.

B. 9ol-brom0-16[3-methyl 1 1 18,17t,21 trihydroxy-LS-pregnadiene-3,20-dione 21-atate.In the manner of Example 10B, the6fi,9ot-dibromo-l,4-pregnadiene of Example 18A, is reacted with zinc inaqueous ethanol and the resulting product isolated and purified in thedescribed manner to give 9OC-bI'OII1Q-16fi-lll6thYl-l1'p,17ot,21-trihydroxy-l,5-pregnadiene-3,20-dione Zl-acetate.

C. 9a bromo 16B methyl 1113,170 21 tl'ihydroxy 1,5 pregnadl'ene 3,20 dione.T he 21-acetate of Example lSB tis convertedto the corresponding21-alcohol with the aid of a culture of Flavobacterl'um dehydrogenans inthe manner of Example 2 to give 9a bromo 165 methyl 11B,l7ot,21trihydroxy 1,5- pregnadiene 3,21) dione. H k p I 7 Alternatively, thecompound of example may be prepared by the alternative procedureofExample 13 from 65,92: dibromo 16 3 methyl 11fl,17ot,21 trihydroxy 1,4pregnadiene 3,20 dione 21 acetate to give 90: bromo 16B methyl11}3,17bc,21 -trihydroxy- 1,5 pregnadiene- 3,20 dione.

"1 1 EXAMPLE 19 a bromo 16a meth l 1 1p,17tt,21 ttih thex 1,5pregnadl'eile 3,20 dione A. 65,9 dibromo 16m meth l 11p,17e,21 thhydroxy 1,5 pregmldiene 3,20 dione acetate;- The requisite intermediate,90c bromo 16a; methyl- 11,B,1706,21 tn'hydroxy 1,4 pregnadiene 3,20-dione 21 acetate is prepared by the process of copending applicationSerial No. 733 843 of Rausser et al., filed May 8, 195 8. l l

The above bromo intermediate is brominated, and the resulting productisolated in the manner of Example 10A to yield 613,90: dibromo 16amethyl 115,1711; 21 trihydroxy 1,4 pregnadiene 3,20 dione 21- acetate.

B. 9OL-l7rOll'lU-160L-m thyl 11fl,17et,21 trihydroxy 1,5- pregnadiene3,20 dion'e 21 aceta-le.-In the manner of Example 10B, the 6/3,9btdibromo 1,4 pregnadienc of Example 19A is reacted With zinc in aqueousethanol to give 90c bromo 16a methyl 11B,17C,21 trihydroxy 1,5pregnadiene 3,20 dione 21 acetate.

C. 904 hrbmo 16a m et hyl J1,8,17ot,21 frillydroxy 1,5 pregnudl'ehe 3,20dione.The 21-acetate pregnadiene of Example 19B is hydrolyzed to'thecorresponding 21-alcohol With the aid of a culture of Flavobacteriumdehydrogenans in the manner of Example 2 to give 9a bromo 16a methyl11fl,17ot,2l -trihydroxy 1,5 pregnadien'e 3,20 dione.

The compound of this example may also be prepared from 613,906 dibromo16a methyl 11B,17et,2l trihydroxy 1,4 pregnadiene 3,20 dione 21 acetate(the compound of Example 19A) by the alternate process of Example 13yielding 9a bromo 16m methyl- 11}3,17ot,2l trihydroxy 1,5 pregnadiene3,20 dione.

EXAMPLE 20 9a bromo a methyl 17tx,21 e dihydroxy 1,5- pregn'allie'ne3,11,20 triohe A. 6,8,9 dibromo 16a meth l 17e,21 eh'h -v droxy 1,4prgnadien 3,11,20 t rione. 21 acetateThe requisite intermediate, 90cbromo 160tmethyl 17ot,21 dihydroxy 1,4 pregnadiene 3,11,- 20 trione 21acetate, is prepared according to the procedure in co-pendingapplication Serial No. 733,843 of Rausser ettaL, filed May 8, 1958.

This intermediate is brominated in the manner of Example 10A and theresulting product isolated in the described manner to give 65,9a dibromo16a methyl- 17ot,21 dihydroxy 1,4 pregnadiene 3,11,20 trione 21-acetate.

B. 90a bromo 160a methyl 170:,21 dihydroxy- ],5 pregm' diene 3,11,20lrl'one 21 acelale.'-The dibromide of Example 20A is reacted with zincin aqueous ethanol in the manner of Example 10B, andthe resultingproduct isolated and purified in the described manner to 'give 904 bromo16a methyl 17ot,'21 dihydroxy 1,5 pregnadiene 3,11,20 trione 21 acetate.

C. 9st bromo 16a methyl 17ot,21 dihydroxy- ],5 pregnadl'ene 3,11,20triona-Jn the manner described in Example 2, the 21-aceta-te of aboveExample 20B is hydrolyzed to the corresponding 21-a1cohol with the aidof a culture of Flavobacterium dehydrogenans to give 9a bromo 1'6 methyl17u,21 dihydroxy- 1,5 pregnadiene 3,11,20 trione.

Alternatively, the compound of this example may be prepared by thealternative process of Example 13 from '6/5,9a dibromo 16a methyll7ot,2l dihydroxy- 1,4 pregnadiene 3,11,20 trione 21 acetate.

EXAMPLE 21 9a bromo 16p methyl 17a,21 dihydroxy 1,5-

pregnadl'ene 3,11,20 z rione A. 65,9ot dibromo 16B methyl 17ot,21dihydroxy 1,4 pregnadiene 3,11,20 trione 21 ace- '15 tate.-The requisiteintermediate, 9a bromo 1618- methyl 1711,21 dihydroxy 1,4 pregnadiene3,11,- 20 trione 21 acetate is prepared by the process of copendingapplication Serial No. 733,843 of Rausser et al., filed May 8, 1958.

The 90: bromopregnadiene intermediate is brominated and isolated in themanner of Example 10A to give 6a,- 900 dibromo 16/3 methyl 1704,21dihydroxy 1,4- pregnadiene 3,11,20 trione 21 acetate.

B. 9a bromo 16B methyl 17oc,21 dihydroxy- 1,5 pregnadiene 3,11,20 trione21 acetate.The dibromide of Example 21A is reacted with zinc powder inaqueous ethanol in the manner of Example 10B, and the resulting productisolated and purified in the described manner to give 90c bromo 16,8methyl 17a,21 dihydroxy 1,5 pregnadiene 3,11,20 trione 21 acetate.

C. 90: bromo 16B methyl 170:,21 dihydroxy- 1,5 pregnadiene 3,11,20trione-The 21 acetate of Example 21B is hydrolyzed to the correspondingalcohol with the aid of a culture of Flavobacterium dehydrogenans toyield 90: bromo 165 methyl 1711,21 dihydroxy 1,5 pregnadiene 3,11,20trione.

EXAMPLE 22 A. 6 3 bromo 16m methyl 11;3,17u,21 trihydroxy 1,4 Lpregnadiene 3,20 dione 21 acetate. The requisite intermediate, 16ccmethyl 115,111,21- trihydroxy 1,4 pregnadiene 3,20 dione 21-acetate, isprepared by the process of co-pending application Serial No. 733,843 ofRausser et al., filed May 8, 1958.

The 160: methyl 1,4 pregnadiene intermediate is brominated and theresulting product isolated in the manner of Example 10A to give '65bromo 16amethyl 11/3,17a,21 trihydroxy 1,4 pregnadiene- 3,20 dione 21acetate.

B. 16oz methyl 11B,17oc,21 trihydroxy 1,5 pregnadiene 3,20 dione 21acetate-The 6,8 bromo- 1,4 pregnadiene of Example 22A (500 mg.) isdissolved in 150 ml.'of absolute ethanol, then 25 ml. of water and g. ofzinc powder are added. The suspension is stirred at room temperature for8 hours, after which time the zinc is filtered. The filtrate isconcentrated in vacuo to a residue which is crystallized from acetone togive 160: methyl 11fl,17oc,21 trihydroxy- 1,5 pregnadiene 3,20 dione 21acetate.

Alternatively, the compound of this example is prepared from the 618bromo 1,4 pregnadiene of Example 22A with zinc in ethanol by theprocedure of Example B.

C. 160: methyl 11B,17ot,21 trihydroxy 1,5 pregnadiene 3,20 dione-The21-acetate of above Example 22B is hydrolyzed to the corresponding21-alcohol with the aid of a culture of Flavobacterium' dehydrogenans inthe manner of Example 2 to give 160: methyl- 11,8,17a,21 trihydroxy 1,5pregnadiene 3,20 dione.

Alternatively, the product of this example may be prepared from 6,8bromo 160a methyl 11B,17u',21- trihydroxy 1,4 pregnadiene 3,20 dione 21acetate by the alternative process of Example 13.

EXAMPLE 23 16,3 methyl 11fii,17a,21 trihydroxy 1,5 pregnaa'iene 3,20dione A. 63 bromo 16B methyl 11fl,17a,21 trihydroxy 1,4 pregnadiene 3,20dione 21 acetate:-- The requisite intermediate, 16/? methyl 11B,17a,21trihydroxy 1,4 pregnadiene 3,20 dione 21 acetate, is prepared accordingto the process of co-pending application Serial No. 733,843 of Rausseret al., filed May 8, 1958.

This 16B-methyl-1,4-pregnadiene intermediate is brominated, and theresulting product isolated and purified.

16 in the manner of Example 10A to give 6B-bromo-16fimethyl 11fl,17a,21trihydroxy 1,4 pregnadiene 3, 20-di0ne ZI-acetate.

B. 16,8 methyl 11/3,17on,21 trihydroxy 1,5 pregnadiene-3,20-di0ne21-acetate.-The 6fl-bromo-L4-pregnadiene of Example 23A is reacted withzinc powder in aqueous ethanol in the manner of Example 22B, and theresulting product isolated and purified in the described manner to give16f3-methyl-11B,17o=,21-trihydroxy-1,5-

pregnadiene-3,20-dione 21-acetate.

C. 165 methyl 11fl,17a,21 trihydroxy 1,5-pregnadiene-3,20-di0ne.--'Ihe21-acetate of Example 23B is hydrolyzed to the corresponding 21-alcoholwith the aid of a culture of F lavobacterium dehydrogenans in the mannerdescribed in Example 2 to give 16B-methyl-11/3,l7u,2l-trihydroxy-l,5-pregnadiene-3,20-dione.

Alternatively, the compound of this example may be prepared from6,8-brom0-16/3-methyl-l1B,17a,21-trihydroxy-l,4-pregnadiene-3,20-dione21-acetate (the compound of Example 23A) by the alternative process ofExample 13.

EXAMPLE 24 16a-methyl-1 7a,21-dihydroxy-1,5-pregnadiene-3,1 1 ,20-

trione A. 6,8 bromo 16a methyl 170:,21 dihydroxy 1,4-pregnadiene-3,11,20-trione 21-acetate.The requisite intermediate, 16amethyl 1701,21 dihydroxy 1,4 pregnadiene-3,11,20-trione 21-acetate, isprepared according to the process of co-pending application Serial No.733,843 of Rausser et al., filed May 8, 1958.

This 16a-meflhyl-l,4-pregnadiene intermediate is brominated and theresulting product isolated in the manner of Example 10A to give6,8-bromo-16a-methyl-17a,21-dihydroxy-l,4-pregnadiene-3,11,20-trioneZI-acetate.

B. methyl 170:,21 dihydroxy 1,5 pregnadiene-3,11,20-trione21-acetate.-The 6fi-bromo-1,4-pregnadiene of Example 24A is reacted withzinc powder in aqueous ethanol in the manner of Example 22B, and theresulting product isolated and purified in the described manner to give16a-methyl-17a,21-dihydroxy-1,5-pregnadiene-3,11,20-trione 21-acetate.

C. 1605 methyl 170;,21 dihydroxy 1,5 pregnadiene-3,11,20-tri0ne.-The21-acetate of Example 24B is hydrolyzed to the corresponding 21-alcoholwith the aid of a culture of Flavobacterium dehydrogenans in the mannerdescribed in Example 2 to give 16a-methyl-17a,21-dihydroxy-1,5-pregnadiene-3,1 1,20-trione.

Alternatively, the compound of this example may be prepared from6;8-bromo16a-methyl17a,2l-dihydroxy-l, 4-pregnadiene-3,11,20-trione21-acetate (the compound of Example 24A) by the alternative process ofExample 13.

EXAMPLE 25 16fl-methyl-1 7a,21 -dihydroxy-1,5-pregnadiene-3,1 1,20-trione A. 6ft bromo 16,8 methyl 170:,21 dihydroxy 1,4-pregnadiene-3,11,20-tri0ne 21-acetate.-The requisite intermediate,16B-methyl-17a,2l-dihydroxy-l,4-pregnadiene-3,11,20-trione 21-acetate isprepared according to the process of co-pending application Serial No.733,843 of Rausser et al., filed May 8, 1958.

This 16/3-methyl-1,4-pregnadiene intermediate is brominated and theresulting product isolated in the manner of Example 10A to give6B-bromo-16fl-methyl-17a,21-dihydroxy-1,4-pregnadiene-3,11,20-trioneZI-acetate.

B. methyl ;,21 dihydroxy 1,5 pregnadiene-3,11,20-tri0ne 21-acetate.-The6fi-bromo-1,4-pregnadiene of Example 25A is reacted with zinc powder inaqueous ethanol in the manner of Example 22B, and the resulting productisolated and purified in the described manner to give16fi-methyl-17a,2l-dihydroxy-l,5-pregnadiene-3,11,20 trione 21-acetate.

C. 16,6 methyl 170:,21 dihydroxy 1,5 pregna- 17diene-3,11,20-trione.-The 21-acetate of Example 25B is hydrolyzed to thecorresponding 21-alcohol with the aid of a culture of Flavobacteriumdehydrogenans in the mannot described in Example 2 to give16fi-methyl-17a,2ldihydroxyl ,5-pregnadiene-3 ,1 1,20-trione.

Alternatively, the compound of this example may be prepared from6B-bromo-16fi-methyl-17u,2l-dihydroxy- 1,4-pregnadiene-3,11,20-trione21-acetate (the compound of Example 25A) by the alternative process ofExample 13.v

EXAMPLE 26 16a-ethyl-1 7ot,21-dihydroxy-1,5-pregnadiene-3,11,20-

, trione A. 613 bromo 161x etihyl 17a,21 dihydroxy 1,4pregnadiene-3,11,20-trine 21-acetate.--The requisite intermediate,l6a-ethyl-170:,2l-dihydroxy-l,4-pregnadiene- 3,11,20-trione 2l-acetateis prepared according to the process of co-pending application SerialNo. 733,843 of Rausser et al., filed May 8, 1958.

This IGm-ethyI-IA- regnadiene intermediate is brominated and theresulting product isolated in the manner of Example 10A to give6B-bromo-16u-ethyl-l7a,21-dihydroxy-1,4-pregnadiene-3,1 1,20-trione 2l-acetate.

B. 160: ethyl .l7oc,21 dihydroxy 1,5 pregnadiene 3,11,20-trione21-acetate.-Thc GB-bromo-lA-pregnadiene of Example 26A is reacted withzinc powder in aqueous ethanol in the mannerof Example 103, and theresulting product isolated and purified in the described manner to give16a-ethy1-l7a,21-dihydroxy-l,5-pregnadiene-3,11,20-trione 21-acetate. v

C. 16a ethyl 17oc,2l dihydroxy 1,5 pregnadiene 3,11,20-tri0ne.The2l-acetate of Example 26B is hydrolyzed to the corresponding 21-alcoholwith the aid of a culture of Flavobacterium dehydrogenans in the mannerdescribed in Example 2 to give16a-ethyl-17a,2l-dihydroxy-1,5-pregnadiene-3,1 1,20-trione.

Alternatively, the compound of this example may be prepared from6/3-bromo-l6a-ethyl-17a,21-dihydroxy-1,4- pregnadiene-3,11,20-trione21-acetate (the compound of Example 26A) by the alternative process ofExample 13.

EXAMPLE 27 16a-ezlhyl-l1B,]7a,21-trihydr0xy-1,5-pregnadiene-3,20- dioneA. 6,8 bromo 16a ethyl 11fl,17a,21 trihydroxy 1,4-pregnadiene-3,20-dione21-acetate.-The requisite intermediate,l6a-ethyl-l1/3,17a,2l-trihydroxy-1,4-pregnadiene-3,20-dione 2l-acetate,is prepared according to the process of co-pending application SerialNo. 733,843 of Rausser et al., filed May 8, 1958.

This 16a-etlhyl-l,4-pregnadiene intermediate is brominated and theresulting product isolated in the manner of Example 10A to give6B-bromo-16a-ethy'l-l1,8,17a,21-trihydnoxy-l,4-pregnadiene-3,20-dione21-acetate.

B. 16a ethyl 11/3,17u,21 trihydroxy 1,5 pregnadiene-3,20-dione21-acetate.-The 6fi-bromo-1,4-pregnadiene of Example 27A is reacted withzinc powder in aqueous ethanol in the manner of Example 10B, and theresulting product isolated and purified in the described manner to give16a-ethyl-11,8,17a,21-trihydroxy-1,5-pregnadiene-3,20-dione ZI-acetate.

C. 16a ethyl 11p,17u,21 trihydroxy 1,5 pregnadiene-3,20-di0ne. -The21-acetate of Example 27B is hydrolyzed to the corresponding 21-alcoholwith the aid of a culture of Flavabacterium dehydrogenans in the mannerdescribed in Example 2 to give16u-ethy1-l1/3,17a,2ltrihydroxy-l,5-pregnadiene-3,20-dione.

Alternatively, the compound of this example may be prepared from 6,8bIOInOl6cc-thYl-1l;9,17u,2l trihydroxy-1,4-pregnadine-3,20-dione21-acetate (the compound of Example 27A) by the alternative process ofEx ample 13.

18 1 EXAMPLE 28 9u-br0mo16a-ethyl-11fL1 7a,21-trihydr0xy-1,5-pregnadiene-3,20-di0ne A. 6,9,9 dibromo 16a ethyl-115,17a,21-trihydroxy-1,4-pregnadieneiZO-dione 21-acetate.- -'Ihe requisite intermediate,9u-br0n10-l6oc-6thYl- 11 B,17oz,21 trihydroxy-1,4-pregnadiene-3,20-dione 21-acetate, is prepared accord-' ing to theprocess of copending application Serial No. 733,843 of Rausser et al.,filed May 8, 1958.

This 9a-bromo-16a-ethyl-l,4 pregnadiene intermediate is brominated andthe resulting product isolated in the manner'of Example 10A to give65,9a-dibromo-l6m-ethyl-1118,17a,21-trihydroxy-1,4-pregnadiene-3,2Odione 21- acetate. 7

B. Qua-bromo 16oz ethyl 115,17a,21-trihydr0xy-1,5-pregnadiene-3,20-di0ne 21-acetate.The 6l3-bromo-l,4- pregnadiene ofExample 28A is reacted with zinc powder in aqueous ethanol in the mannerof Example 10B, and the resulting product isolated and purified in thedescribed manner to give9a-bromo-16a-ethyl-11B,l7a,21-trihydroxy-1,5-pregnadiene-3,20-dione21-acetate.

C; 90: bromo 16a ethyl 1113,]7a,21-trihydr0xy-1,5-pregnadiene-3,20-di0ne.-Th'e 2l-acetate of Example 28B is hydrolyzed tothe corresponding 21-alcohol with the aid of a culture of Flavobacteriumdehydrogenans in the manner described in Example 2 to give9ot-bIOII10-16ocethyl-11B,17,2l-trihydroxy-1,5-pregnadiene-3,20-dione.

Alternatively, the compound of this example may be prepared from6fl,9a-dibromo-l6a-ethyl-l1fi,17a,21 trihydroxy 1,4 pregnadiene 3,20dione 21-acetate (the compound of Example 28A) by the alternativeprocess of Example 13.

EXAMPLE 29 9a-flu0r0-16a-ethyl-11,8,17a,21-trihydr0xy-1,5-pregnadiene-3,20-dione 1,5 pregnadiene 3,20 dione 21-acetate.-The 6flbr0mo-9a-fluoro-1,4-pregnadiene of Example 29A is reacted with zinc powderin aqueous ethanol in the manner of Example 10B, and the resultingproduct isolated and purified in the describedrmanner to give9oc-flllOIO-l6oc-ethYl- 11,6,l7ot,2l-trihydroxy-l,S- regnadiene 3,20dione 21- acetate.

C. 90: fluoro 16a ethyl 11,6,17a,21 trihydroxy- 1,5 pregnadiene 3,20di0ne.-The 2l-acetate of Example 29B is hydrolyzed to the corresponding21-alcohol with the aid of a culture of Flavobacterium dehydrogenans inthe manner described in Example 2 to give9afluoro-16ot-ethyl-1lfl,17a,2l-trihydroxy-l,5 pregnadiene- 3,20-dione.

Alternatively, the compound of this example may be prepared from 65bromo 90c fiuoro 16a ethyl l1B,17a,21 trihydroxy 1,4 pregnadiene 3,20dione 2l-acetate (the compound of Example 29A) by the alternativeprocess of Example 13.

EXAMPLE 3O 9oc-flu0r0-16a-ethyl-1 711,21-dihydroxy-1,S-pregnadiene- 3,11,ZO-trione "*19 pregnadiene-3,11,20-trione 21-acetate, is preparedaccording to the process of co-pending application Serial No. 733,843 ofRausser et al., filed May 8, 1958.

This 9u-fluoro-16a-ethyl-l,4-pregnadiene intermediate is brominated andthe resulting product isolated in the manner of Example 10A to give6B-bromo9a-fluoro-16aethyl-17a,21-dihydroxy 1,4 pregnadiene 3,11,20-trione ZI-acetate.

B. 9m fluoro 16cc ethyl 170:,21 dihydroxy 1,5- pregnadiene 3,11,20trione 21-acetate.-The 6fl-bromo- 9a-fluoro-1,4-pregnadiene of Example30A is reacted with zinc powder in aqueous ethanol in the manner ofExample 103, and the resulting product isolated and purified in thedescribed manner to give9a-fluoro-16a-ethyl-17a,2ldihydroxy-1,5-pregnadiene-3,11,20-trione21-acetate.

C. 9a fluoro 16a ethyl I7u,21 dihydroxy 1,5-pregnadiene-3,11,20-trione.The 21-acetate of Example 30B is hydrolyzedto the corresponding 21-alcohol with the aid of a culture of Flavobacterium dehydrogenans in the manner described in Example 2 to give90L-flllOl'O-16OL- ethyl-17a,21-dihydroxy-1,5-pregnadiene-3 ,11,20-trione.

Alternatively, the compound of this example may be prepared from6e-bromo-9a-fluoro-16u-ethy1-17a,21 dihydroxy 1,4 pregnadiene 3,11,20trione 21-acetate (the compound of Example 30A) by the alternativeprocess of Example 3.

EXAMPLE 31 16oz-n-butyl-1 711,21 -dihydroxy-1 ,S-pregnadiene-3,11,20-trine A. 66 bromo 16a n butyl 17a,21-dihydr0xy-1,4-pregnadiene-3,11,20-tri0ne 21-acetate.-The requisite intermediate,16a-n-butyl-17u,21 dihydroxy 1,4 pregnadiene-3,11,20-trione 21-acetate,is derived from 16a-nbutylcortisone 21-acetate, which, in turn, isprepared according to the process of copending application Serial No.733,843 of Rausser et al., filed May 8, 1958. 160a n butylcortisone21-acetate is subjected to the action of a culture of Corynebacteriumsimplex (A.T.C.C. 6946) in the manner disclosed in US. Patent No.2,837,464, and the resulting reaction mixture extracted with chloroformin the described manner. The chloroform extracts are combined andconcentrated to a residue which, after crystallization from acetone,gives 16a-nbutyl-17a,21-dihydroxy-1,4 pregnadiene 3,11,20 trione21-acetate.

16a n butyl 17a,21 dihydroxy 1,4-pregnadiene- 3,11,20-trione 21-acetateis brominated and the resulting product isolated in the manner ofExample 10A to give 6fi-bromo-16a-n-butyl 1702,21 dihydroxy1,4-pregnadiene-3,11,20-trione ZI-acetate.

B. 16a n butyl 1711,21 dihydroxy-1,S- regnadiene- 3,11,20-trione 21acetate.-The 6l3-bromo-l6a-n-butyl- 1,4-pregnadiene of above Example 31Aisreacted with zinc in aqueous ethanol in the manner of Example 10B andthe resulting product isolated and purified in the de scribed manner togive 16u-n-butyl-17a,21-dihydroxy-l,5 pregnadiene-3, 1 1,20-trione21-acetate.

C. 16oz nbutyl 17a,21 dihydroxy-1,S-pregnadiene- 3,11,20-trione.The21-acetate of above Example 31B is hydrolyzed to the corresponding21-alcohol with the aid of a culture of F lavobacterium dehydrogenans bythe procedure described in Example 2.

Alternatively, the compound of this example may be prepared from6fl-bromo-l6a-n-butyl-Hull-dihydroxy- 1,4-pregnadiene-3,11,20-trione21-acetate (the compound of Example 31B) by the alternative process ofExample 13.

EXAMPLE 32 16,8-n-butyl-17a,21-dihydroxy-1,S-pregnadiene- 3,11,ZO-trione The requisite intermediate,16fl-n-butyl-17a,21-dihydroxy-1,4-pregnadiene-3,l1,20-trione 21-acetate,is derived from 16B-n-butylcortisone 21-acetate which is prepared bytheprocess ofco-pendingapplication Serial No; 733,843 of Rausser et a1,filed May 8, 1958. When subjected to the action of a culture ofCorynebacterium simplex in the manner disclosed in US. Patent No.2,837,464, and the resulting product isolated and purified as describedin Example 31A, 16fl-n-butylcortisone' 21- acetate is converted to16l3-n-butyl-l7a,21-dihydroxy-1,4- pregnadiene-3,11,20-trione21-acetate.

In the manner of Example 31,16B-n-butyl-17a,21-dihydroxy-1,4-pregnadiene-3,11,20 trione 21 acetateis brominated to the corresponding 6/3-bromo-derivative which, in turn,is reacted with zinc in aqueous ethanol to give16,8-n-butyl-17a,21-hydroxy-1,5-pregnadiene-3,11,20- trione 21-acetate.

The 21-acetate is then hydrolyzed to the 2l-alcohol in thedescribed'manner to givel6fl-n-butyl-17a,21-dihydroxy-'1,S-pregnadiene-3 ,1 1,20-trione.

EXAMPLE 3'3 To one gram of16oz-methyl-17a,2l-dihydroxy-1,5-pregnadiene-3,11,20-trione, thecompound of Example 24, there is added 0.5 ml. of n-propionic anhydridein 2.3 ml. of pyridine. After standing for one hour at room temperature,the mixture is poured into ice and hydrochloric acid. The resultingprecipitate is filtered and recrystallized from aqueous methanol toyield 16x-methyl- 17a,2l-dihydroxy-LS-pregnadiene 3,11,20 trione21-npropionate.

Similarly, any one of the 21-hydroxy-1,5-pregnadiene compounds of ourinvention may be treated with propionic anhydridein the above describedmanner to yield the corresponding 21-n-propionate ester.

By substituting anhydrides of other lower alkanoic acids, such asacetic, butyric, valeric and the like, for propionic anhydride in theprocedure of Example 33, there may be obtained the corresponding21-lower alkanoate of any 16-alkyl-1,5-pregnadiene-2l-hydroxy startingcompound.

EXAMPLE 34 16a-methyl-1 70:,21dihydroxy,1,5-pregnadiene- 3,11,20-tri0neG. Difco yeast extract 3-10 Corn steep liquor 1 Dextrose 10 Distilled ortap water, q.s. to one liter.

Each flask is inoculated with spores from an agar medium culture ofChaetomium funicolum (QM No. 33C) or with a 110% submerged inoculumwhich has been grown for 24-48 hours. The mixture is incubated byshaking the flasks on a rotary shaker for 24-48 hours at 28 C. atapproximately 250 rpm. To each of the flasks (now showing prolificgrowth) is added, aseptically, 50 mg. of16u-methyl-17a,21-dihydroxy-1,4-pregnadiene-3,1 1,20- trione in 2 ml. ofethanol. The fermentation mixture is then incubated and shaken for 24-72hours at 28 C. after which time complete conversion occurs. The mixtureis extracted thoroughly with chloroform and the small fraction of theproduct which is retained within the mycelium is extracted by boilingthe mycelium in chloroform for a few minutes. The chloroform extractsare combined and evaporated to a residue yielding approximately 5 mg. ofcrude material. The residue is triturated with methanol aifording acrystalline solidfwhich. is purifiedby crystallization from acetoneyielding 16a-methyl-6,8,.17ci,2I tnihydroxy1 ,4-pregnadiene-3,.l1,2ltltri'one. T I

B. 16a-methyl-6fl,17a,2l irihydroxy-I,4-pregnadiene- 3,11,2'0-tri0ne21-acetate.-A solution of 25 mg. of the compound of Example 34A in. 2m1. of. anhydrous pyridine ispoured into 6 mlof acetic anhydride in: anitrogen atmosphere. The mixture is stirred for 30 minutesand thenpoured into dilute sulfuric acid and ice. The resultant precipitate isremoved by filtration, dried. and'crystallizedfrom methanol, yieldingl6a-metl'1yl-6B,17a,2l.-trihydroxy-1,4-pregnadiene-3 ,1 1,20-trione '2l-ac'etatei C. 16a-methyl-6/9,17u,21 tiihydroxy-l,4-pregnadine-3,11,20-trine 6-methzmesulfonate '2'1-acetate.-l0O- mg. of16u-methyl-6B,1 7a,21-trihydroxy-1,4-pregnadienee3',1 1, 20-trione2l-acetate, prepared as Example 34B, is dissolved in 0.4 ml; ofmethylene chloride and0l4 ml. of pyridine. The solution is cooled to 0C., 1.6;ml. of methanesulfonyl chloride added, and the mixture stirredfor 2 hours at 0 C., thenleft overnight at room temperature. Thereaction mixture is then poured into Water, the resulting precipitatewhich is filtered and air driedis 16wmethyl.-6B;l7a,21-trihydroxy-1,4-pregnadiene 3,11, 20-trione 6-methanesulfonate2'1-acetate'.

D. 160L-m8thyl 17a,2l dihydroxy LS-pregnadiene- 3,11,20-trione2-1-ace'tate.-The 6-niethanesulfonate-l 6amethyl-1,4-pregnadiene (55mg.) of above Example 34C- of substantially 16a-methyl-l7bz,21-dihydroxy-l,5-pregnadiene-3,l1,20-trione Zl-acetate.

E. Mix-methyl 1 7:1,2] dihydroxy-I,5-pregnadiene-3, 11,20-tri0ne.-The2l-acetate ofiExample 34D is hydrolyzed to the corresponding 21-alcoholin the manner described in Example 2 to givel6a-methyl-l7a,21-dihydroxy-1,5-pregnadiene-3,l 1,20-trione.

Alternatively, the compound of this example may be prepared as describedin following procedures F and G.

F. 16a-methyl 6fl,17a,21 trihydroxy-Z,4-pregnadiene- 3,11,20-tri0ne6,8,21-diacetate.'A solution of 25 mg. of the compound of Example 34A in2 ml. of anhydrous pyridine is poured onto 100 mg. of acetic anhydridein an anhydrous atmosphere. The mixture is stirred for 30 minutes andthen poured into dilute sulfuric acid and ice. The resultant precipitateis removed by filtration, dried and crystallized from methanol, yieldingl6a-methyl-6fi, 17a,21-trihydroxy-1,4-pregnadiene 3,11,20 trione 65,21-diacetate.

G. 16a-methyl-17a,21 a'ilzydroxy 1,5-pregnadiene-3, 11,20-tri0ne21-acetate.The 65,2l-diacetate of Example 34F is reacted with zinc andalcohol in the manner described in Example 34D to givel62x-methyl-17a,21-dihydroxy1,5-pregnadiene-3, 1 1,20-trione 2l-acetate.

The 21-acetate of this example is. converted to the corresponding21-hydroxy-1,5-pregnadiene in the manner of Example 2.

A third alternative process is described infollowing procedures H and I.

H. 1600 methyl '6/3,17a,2l trihydroxy 1,4 pregna-l dime 3,11 ,20 trione6,3 p toluenesulfonate 21 acetate.A solution of 100 mg. of the productof Example 34A in 2 ml. of anhydrous pyridine is treated with 100 mg. ofp-toluenesulfonyl chloride and the mixture is stirred for 1 hour. Thesolution is poured into aqueous sulfuric acid and ice, and the resultingprecipitate is removed by filtration, washed with water, dried, andcrystallized from acetone-hexane to give 16u-methyl-6B,17a,21-trihydroxy-l,4-pregnadiene-3,11,20-trione 6 8-p-toluenesulfonate21-acetate.

I. 16a methyl 17,2ldihydroxy 1,5 pregnadiene 3,11,20 trion'e '21acetate.--'Ihe 6-p-toluenesulfonate 21-acetate 1,4-pregnadiene of aboveExample 34H is reacted with zinc powder in the manner described.

22 in Example 22B to give 16a-methyl=17u,21-dil1ydroxy-1,5-pregnadiene-3,1 1,20-trione 21-acetate.

The 21-acetate of the lfia-methyl-1,5-pregnadiene prepared above 'ishydrolyzed to the corresponding 21-hydroxy compound in the mannerdescribed in Example 2.

EXAMPLE 35 9ot-brom0-16a-ethyl-17oz,21-dihydroxy-1,5-pregnadiene-3,1 1,20-tri0ne A. bromo 16a ethyl 176:,21 dihydroxy -1,4-

pi'egnadiene 13,11,20 trione 21 acetate-The requi site intermediate,9a-bromo-16a-ethyl-1 1fi,17a,2;1.-trihydroxy-l,4-pregnadiene-3,20-dione21-acetate, is prepared as described in co-pending application SerialNo.- 7 33,843 of Rausser et al., filed May 8, 1958.

To a solution of 0.3 g. of 90t-bI'OIIl0-160L-ethy1-170t,21-dihydroxy-1,S-pregnadiene-3,20-dione 21-acetate in 15 ml. of aceticacid, there is added dropwise a solution of 60 mg. of chromium trioxidein 1 ml. of water and 3 ml. of acetic acid. The resultant mixture isallowed to stand for 5 hours, then is dilutedwith water, and extractedwith methylene chloride; The organic extracts are washed with water,dried over magnesium sulfate, filtered and evaporated to a residue whichis crystallized from methanol to give9u-bromo-16u-ethyl-17a,21-dihydroxy-l,4-pregnadiene-3,11,20-trione21-acetate.

1,,4 pregnad iene 3,11,20 triqrge "21 actate.-The 9bebromo-16aethyl-1,4-pregnadiene of Example 35A is brominated and the resultantproduct isolated in the manner described iii-Example 10A to give 63,9a-dibromo-16alayer is separated from the aqueous layer. The chlo1'0,

form'solutionis concentrated in vacuo to yield 9a-bromo- 16m ethyl170;,21 dihydroxy 1,5 pregnadiene 3,11, 20-trione ZI-acetate. p V v D.9a bromo 16a ethyl 170:,21 dihydroxy 1,5- pregnadiene 3,1],20-triona-The21-acetate of Example 35C is hydrolyzed to the corresponding 2l-alcoholwith the aid of a culture of Flavobacterium dehydrogenans to yield 90!.bromo a ethyl 1711,21 dihydroxy 1,5- pregnadiene-3 ,1 1,20-trione.

We claim: 1. 1,5-pregnadienes having the following formula:

CHzOR lower alkyl; and Ris a m'einberof the group consisting g of H andlower alkanoyl.

ethyl-17a,21-dihydroxy-l,4-pregnadiene-3,1 1,20-trione 21- 23 2,1,5-pregnadien'es having the formula:

a CH3 ornoR wherein X is a halogen of atomic weight less than 126. *3.The 21-lower alkanoyl esters of the compounds of claim 2.

4. 1,5-pregnadienes having the formula:

wherein X is a halogen of atomic weight less than 126 and Z is loweralkyl.

'5. The 21-lower alkanoyl esters of the compounds of claim 4.

6. 1,5-pregnadienes having the formula:

wherein X is a halogen of atomic weight less than 126.

7. The 21-lower alkanoyl esters of the compounds of claim 6.

8. 1,5-pregnadienes having the formula:

- trihydroxy 1,5 pregnadiene-3,20-dione. r

24 16. 9oz fluoro 170:,21 dihydroxy 1,5 1 pregnadiene 3,11,20-trione.17. 9a fluoro 11B,17a,21 trihydroxy 1,5 pregnaa diene-3,20-dione. Y

18. a fluoro 16a methyl 17,21 dihydroxy-. 1,5-pregnadiene-3,11,20-trione. 19. 9a-fiuoro-lfip methyl-lhfll-dihydroxy 1,5 -preg-vnadiene-3,1 1,20-trione.

pregnadiene-3,20-dione.

21. 9a-fluoro-l6B-imethy1 11,9,17a,21 trihydroxy-l,5-pregnadiene-3,20-dione.

22. In the process for the manufacture of 3-keto-A steroids the stepwhich comprises reacting a 3-keto-A -1 steroid having also at the6-position a member of the group consisting of halogen having an atomicweight greater than 19, lower alkanoyloxy, lower a'lkylsulfonyloxy, andphenylsulfonoyloxy with a metal reagent of the group consisting of zincin a lower fatty alcohol, and magnesium in a solvent of the groupconsisting of tetrahydrofuran and lower dialkyl ethers.

23. In the process for the manufacture of a 3-keto-1,5- pregnadiene, thestep which comprises reacting a 3-keto- 6-bromo-1,4-pregnadiene withzinc in a lower fatty a1- cohol.

24. In-the process for the manufacture of pregnadienes.

of the formula:

wherein X is a member of the group consisting of hydrogen and halogenhaving an atomic weight less than 126; Y is a member of the groupconsisting of O and (H,;3OH); Z is a member of the group consisting ofhydrogen and lower alkyl; and R is a member of the group consisting of Hand lower alkanoyl; the step which comprises reacting a compound of theformula:

wherein X, Z and R are as defined above; Y is a member of the groupconsisting of O and (I-LBOR); and A is a member of the group consistingof halogen having an atomic weight greater than 19, lower alkanoyloxy,

lower 'alkylsulfonyloxy, and phenylsulfonyloxy; with a metal reagent ofthe group consisting of zinc in a lower I fatty alcohol .and magnesiumin a solvent of the group consisting of, tetrahydrofuran and lowerdialkyl ethers, and isolating the 1,5-pregnadiene thereby formed.

25. The process of claim 24 including the step of hydrolyzing estergroups present in the 1,5-pregnadiene.

26. In the process for the manufacture of17,2l-dihydroxy-l,5-pregnadiene-3,11,20-trione and the 2l-acetatethereof, the step which comprises reacting 6a,'17a,2l-

trihydroxy-l,4-pregnadiene-3,1l,20-trione' 6,21-diacetate with zinc inalcohol Within a temperature range of about- 50' to C.

26 trione 6,21-diacetate with zinc in alcohol within a temperature rangeof 50 to 100 C.

29. In the process for the manufacture of 16-methy1-1lfl,17a,21-trihydroxy-1,5-pregnadiene-3,20-dione and the ZI-acetatethereof, the step which comprises heating 16- methyl-6,8, 11/i,-17m,21-tetrahydroxy-1,4pregnadiene-3,20- dione 6,21-diacetate withzinc in alcohol Within a temperature range of 50 to 100 C.

No references cited.

UNITED STATES PATENT OFFICE Certificate of Correction Patent No.2,908,696 October 13, 1959 Alexander L. Nussbaum et a1.

It is hereby certified that error appears in the printed specificationof the above numbered satent requiring correction and that the saidLetters Patent should read as correete below. V

Column 1, line l0, for 2-esters read 21-esters; column 4:, lines 22 to3d, the structural formulae of Compounds 11 and III should read as shownbelow instead of as in the patent:

CH: CH:

l mo w 3.0

Hydtohalie 4)" H6 Halogen II III Signed and sealed this 7th day of June1960.

[SEAL] Azjfestz'ng Oficey, Qonwng'gsz'omr of Patents

1. 1,5-PREGNADIENES HAVING THE FOLLOWING FORMULA: